Differences in the signaling pathways of α(1A)- and α(1B)-adrenoceptors are related to different endosomal targeting.
AIMS: To compare the constitutive and agonist-dependent endosomal trafficking of α(1A)- and α(1B)-adrenoceptors (ARs) and to establish if the internalization pattern determines the signaling pathways of each subtype. METHODS: Using CypHer5 technology and VSV-G epitope tagged α(1A)- and α(1B)-ARs sta...
| Main Authors: | , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3663791?pdf=render |
| _version_ | 1818553267784777728 |
|---|---|
| author | Vanessa Segura Miguel Pérez-Aso Fermí Montó Elena Carceller María Antonia Noguera John Pediani Graeme Milligan Ian Christie McGrath Pilar D'Ocon |
| author_facet | Vanessa Segura Miguel Pérez-Aso Fermí Montó Elena Carceller María Antonia Noguera John Pediani Graeme Milligan Ian Christie McGrath Pilar D'Ocon |
| author_sort | Vanessa Segura |
| collection | DOAJ |
| description | AIMS: To compare the constitutive and agonist-dependent endosomal trafficking of α(1A)- and α(1B)-adrenoceptors (ARs) and to establish if the internalization pattern determines the signaling pathways of each subtype. METHODS: Using CypHer5 technology and VSV-G epitope tagged α(1A)- and α(1B)-ARs stably and transiently expressed in HEK 293 cells, we analyzed by confocal microscopy the constitutive and agonist-induced internalization of each subtype, and the temporal relationship between agonist induced internalization and the increase in intracellular calcium (determined by FLUO-3 flouorescence), or the phosphorylation of ERK1/2 and p38 MAP kinases (determined by Western blot). RESULTS AND CONCLUSIONS: Constitutive as well as agonist-induced trafficking of α(1A) and α(1B) ARs maintain two different endosomal pools of receptors: one located close to the plasma membrane and the other deeper into the cytosol. Each subtype exhibited specific characteristics of internalization and distribution between these pools that determines their signaling pathways: α(1A)-ARs, when located in the plasma membrane, signal through calcium and ERK1/2 pathways but, when translocated to deeper endosomes, through a mechanism sensitive to β-arrestin and concanavalin A, continue signaling through ERK1/2 and also activate the p38 pathway. α(1B)-ARs signal through calcium and ERK1/2 only when located in the membrane and the signals disappear after endocytosis and by disruption of the membrane lipid rafts by methyl-β-cyclodextrin. |
| first_indexed | 2024-12-12T09:23:41Z |
| format | Article |
| id | doaj.art-18a98db5f7dc479da6aaa518089cf63d |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-12T09:23:41Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-18a98db5f7dc479da6aaa518089cf63d2022-12-22T00:29:06ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0185e6499610.1371/journal.pone.0064996Differences in the signaling pathways of α(1A)- and α(1B)-adrenoceptors are related to different endosomal targeting.Vanessa SeguraMiguel Pérez-AsoFermí MontóElena CarcellerMaría Antonia NogueraJohn PedianiGraeme MilliganIan Christie McGrathPilar D'OconAIMS: To compare the constitutive and agonist-dependent endosomal trafficking of α(1A)- and α(1B)-adrenoceptors (ARs) and to establish if the internalization pattern determines the signaling pathways of each subtype. METHODS: Using CypHer5 technology and VSV-G epitope tagged α(1A)- and α(1B)-ARs stably and transiently expressed in HEK 293 cells, we analyzed by confocal microscopy the constitutive and agonist-induced internalization of each subtype, and the temporal relationship between agonist induced internalization and the increase in intracellular calcium (determined by FLUO-3 flouorescence), or the phosphorylation of ERK1/2 and p38 MAP kinases (determined by Western blot). RESULTS AND CONCLUSIONS: Constitutive as well as agonist-induced trafficking of α(1A) and α(1B) ARs maintain two different endosomal pools of receptors: one located close to the plasma membrane and the other deeper into the cytosol. Each subtype exhibited specific characteristics of internalization and distribution between these pools that determines their signaling pathways: α(1A)-ARs, when located in the plasma membrane, signal through calcium and ERK1/2 pathways but, when translocated to deeper endosomes, through a mechanism sensitive to β-arrestin and concanavalin A, continue signaling through ERK1/2 and also activate the p38 pathway. α(1B)-ARs signal through calcium and ERK1/2 only when located in the membrane and the signals disappear after endocytosis and by disruption of the membrane lipid rafts by methyl-β-cyclodextrin.http://europepmc.org/articles/PMC3663791?pdf=render |
| spellingShingle | Vanessa Segura Miguel Pérez-Aso Fermí Montó Elena Carceller María Antonia Noguera John Pediani Graeme Milligan Ian Christie McGrath Pilar D'Ocon Differences in the signaling pathways of α(1A)- and α(1B)-adrenoceptors are related to different endosomal targeting. PLoS ONE |
| title | Differences in the signaling pathways of α(1A)- and α(1B)-adrenoceptors are related to different endosomal targeting. |
| title_full | Differences in the signaling pathways of α(1A)- and α(1B)-adrenoceptors are related to different endosomal targeting. |
| title_fullStr | Differences in the signaling pathways of α(1A)- and α(1B)-adrenoceptors are related to different endosomal targeting. |
| title_full_unstemmed | Differences in the signaling pathways of α(1A)- and α(1B)-adrenoceptors are related to different endosomal targeting. |
| title_short | Differences in the signaling pathways of α(1A)- and α(1B)-adrenoceptors are related to different endosomal targeting. |
| title_sort | differences in the signaling pathways of α 1a and α 1b adrenoceptors are related to different endosomal targeting |
| url | http://europepmc.org/articles/PMC3663791?pdf=render |
| work_keys_str_mv | AT vanessasegura differencesinthesignalingpathwaysofa1aanda1badrenoceptorsarerelatedtodifferentendosomaltargeting AT miguelperezaso differencesinthesignalingpathwaysofa1aanda1badrenoceptorsarerelatedtodifferentendosomaltargeting AT fermimonto differencesinthesignalingpathwaysofa1aanda1badrenoceptorsarerelatedtodifferentendosomaltargeting AT elenacarceller differencesinthesignalingpathwaysofa1aanda1badrenoceptorsarerelatedtodifferentendosomaltargeting AT mariaantonianoguera differencesinthesignalingpathwaysofa1aanda1badrenoceptorsarerelatedtodifferentendosomaltargeting AT johnpediani differencesinthesignalingpathwaysofa1aanda1badrenoceptorsarerelatedtodifferentendosomaltargeting AT graememilligan differencesinthesignalingpathwaysofa1aanda1badrenoceptorsarerelatedtodifferentendosomaltargeting AT ianchristiemcgrath differencesinthesignalingpathwaysofa1aanda1badrenoceptorsarerelatedtodifferentendosomaltargeting AT pilardocon differencesinthesignalingpathwaysofa1aanda1badrenoceptorsarerelatedtodifferentendosomaltargeting |